Compositions and Methods of Their Use for Improving the Condition and Appearance of Skin

ABSTRACT

The present invention relates to compositions and methods for treating, preventing and improving the condition and aesthetic appearance of skin, particularly, treating, preventing, ameliorating, reducing and/or eliminating fine lines and/or wrinkles of skin, where the compositions include natural plant constituents, or synthetic equivalents thereof, which inhibit or decrease expression of C-reactive protein expression, production or secretion, thereby reducing, inhibiting, or ameliorating dermatological signs of aging. The compositions of the invention may be topically applied to the skin, or are delivered by directed means to a site in need thereof, in an amount effective in improving the condition and aesthetic appearance of skin.

RELATED APPLICATIONS

This is a divisional application that claims priority to, and thebenefits of, co-pending U.S. patent application Ser. No. 11/647,648,filed Dec. 29, 2006 and incorporated herein by reference.

FIELD OF THE INVENTION

This invention generally relates to cosmetic, dermatological, andpharmaceutical compositions and their use, and more particularly tocosmetic compositions and to their use in improving the condition andappearance of skin.

BACKGROUND OF THE INVENTION

There is an increasing demand in the cosmetics industry to developproducts that may be applied topically to the skin that improve thecondition and appearance of skin. Consumers are interested in mitigatingor delaying the dermatological signs of chronologically orhormonally-aged or photo-aged skin, such as fine lines, wrinkles,drying, and sagging skin, and other conditions due to a progressivedegradation of the skin matrix. During the aging process, the complexionof the skin, i.e., the color and appearance of the skin, deterioratesslowly from aging and/or exposure to sunlight. Numerous cosmetic andmedical treatments have been developed in an attempt to treat aging oraged skin. Such cosmetics or treatments commonly contain organic acidsas their active ingredients or components. These actives are frequentlyassociated with consumer discomfort, such as burning, itching, andredness.

C-Reactive Protein (CRP) is a member of a class of acute phase reactantswhich increases during inflammatory processes, and thereby acts as amarker of inflammation. Measuring and monitoring C-reactive proteinvalues are useful in determining disease progression or theeffectiveness of treatments. Homo sapiens C-reactive protein (CRP) is apentameric protein that has also been found to play a role in heartdisease and is an indicator of risk for hypertension, recurring coronaryevents, and cardiovascular disease (mRNA: Accession No. NM_(—)00567;gene: Accession No. AF442818; mapped to chromosome 1 region q21 to q25;protein: Accession No. AAL48218). Human CRP is composed of five 206amino acid subunits non-covalently bound, each having a molecular weightof 23,017 daltons (Mullenix and Mortensen, Mol. Immunol., 31(8):615-22,1994). Human CRP is normally present in trace amounts in serum, at verylow levels, e.g., 0.8-3 g/mL. However, during infection andinflammation, human CRP levels can increase by 1,000-fold in response tospecific cytokines.

There remains a general need in the cosmetics industry for products thatretard or counter the aging effects on the skin, and more specificallyfor products that produce such effects without undesirable side effects.In particular, there remains a need for topically applied cosmeticcompositions that have anti-aging, anti-inflammatory, and skin texturebenefits using natural plant materials as active components.

The genus Humulus of the Cannabaceae family, have been employed for avariety of purposes. The hop (Humulus) is a small genus of floweringplants, native to the temperate Northern Hemisphere. The female flowers,commonly called hops, are commonly used as flavoring and stabilizersduring the process of brewing beer. Humulene and lupulene are medicallyactive ingredients in hops. Dried female buds having a highmethylbutenol content, are mildly sedating on the central nervoussystem. Hops may be used in the treatment for insomnia, stress, anxiety.Isolated from hops, Xanthohumol, has been shown to have anti-cancerproperties. Xanthohumol has been shown to be toxic to human breastcancer, colon and ovarian cancer cells, as well as has some activityagainst prostate cancer. In addition to its anti-cancer effects, hopshas antibacterial characteristics which stimulate gastric juiceproduction. There are several species (and varieties) belonging to thisgenus, including: H. aculealus, H. americanus, H. cordifolius, H.japonicus (H. japonicas var. variegatus), H. lupulus (H. lupulus subsp.americanus, H. lupulus var. cordifolius, H. lupulus var. lupuloides, H.lupulus var. lupulus, H. lupulus var. neomexicanus, H. lupulus var.pubescens), H. neomexicanus, H. scandens (H. scandens var. varigatus),and H. yunnanensis.

The Humulus scandens, Humulus japonicas, Humulus lupulus, and Humulusyunnanensis species in the genus Humulus are all herbaceous vines withhooked climbing hairs and twine clockwise up. These species generallyare dioecious, i.e., sexually distinct plants; however, individualmonoecious plants, i.e., hermaphroditic, have been identified. Althoughmale and female plants are easily distinguishable during flowering,there are no other morphological differences with which to identify thesex of a plant.

Humulus japonicus, also synonymous with Humulus scandens, is commonlyknown as Asian Hop. Humulus japonicas is a dioecious annual originatingin Eastern Asia. It generally has leaves with 5-7 lobes, longinternodes, and extremely strong hooked climbing hairs. It may be usedas an ornamental primarily for hedges, but its aggressive nature andparticularly strong climbing hairs make it unpleasant to handle and apotentially invasive weed. Because its female inflorescence is smallerthan the cone of H. lupulus and there are few glands on either itsleaves or cones, there is little value for H. japonicus in the brewingindustry.

Humulus lupulus is a perennial that grows again each spring from therhizomes of an underground rootstock. In the wild, H. lupulus spreadsthrough underground rhizomes and by seed. H. lupulus has been separatedinto five taxonomic varieties based primarily on leaf characteristics,including but not limited to, pubescence, leaf hairs, leaf lobes and thenumber of glands.

Brewers have determined that European hops are milder and more aromaticthan those from North America. European-type hops are less pubescent,have fewer glands, and have heart-shaped leaves with few lobes. NativeNorth American hops are generally heavily pubescent and have deeplylobed leaves with five or seven lobes. North American hops have beendescribed as having high resin content and as rich with glands and“buttery” to touch.

Humulus yunnanensis is a dioecious perennial with leaves that are lesslobed and cones larger than those of H. japonicus. H. yunnanensis isdistinguishable by the stiffness of the climbing hairs, pubescent leafsurfaces, small leaf glands, distribution of the trichomes, and largepollen grains. H. yunnanensis has only a few glands on the bracteoles ofthe cone and similar to H. japonicus, has no value for brewing.

It would be desirable to have a composition, preferably topical,comprising a natural plant material to improve the aesthetic appearanceof skin. It would also be desirable to have an effective treatment andpreventative for the dermatological signs of aging, includinginflammation. Therefore, safe, effective and new compositions to treat,prevent, reduce, inhibit, and/or improve the dermatological signs ofaging, would be advantageous for the formulation of treatments andproducts for the skin. As described herein, novel and beneficial methodsand compositions, as well as their mode of action, for the treatment ofwrinkles and the like, as well as for personal care products for theskin, are provided by the present invention.

SUMMARY OF THE INVENTION

It is an object of the present invention to provide a composition thatdelivers an effective level of an active component from a natural plantmaterial that inhibits and decreases the production and/or secretion ofC-reactive protein.

It is another object of the present invention to provide a compositionhaving a natural plant material, blends of natural plant materials, orsynthetic equivalents, in a cosmetically, dermatologically,physiologically, or pharmaceutically acceptable vehicle.

It is a further object of the present invention to provide a compositionhaving a natural plant material, component therefrom, or syntheticequivalent, that results in decreased or inhibited expression ofC-reactive protein thereby reducing dermatological signs ofchronologically or hormonally-aged or photo-aged skin, such as finelines, wrinkles, and sagging skin, where the composition is a cosmetic,dermatological, or pharmaceutical product.

It is yet another embodiment of the invention to provide a method oftreating, preventing, and/or ameliorating the appearance of fine linesand/or wrinkles, comprising applying to skin a composition having anatural plant material, combinations of natural plant materials, orsynthetic equivalents thereof, in an amount effective to prevent,ameliorate and/or reduce dermatological signs of chronologically orhormonally-aged or photo-aged skin, such as fine lines, wrinkles, andsagging skin.

It is still a further object of the present invention to provide acomposition having one or more of the following natural plant materials:H. scandens, H. japonicus, H. lupulus, and H. yunnanensis, in an amounteffective to treat, prevent, control, ameliorate, inhibit, and/or reducedermatological signs of chronologically or hormonally-aged or photo-agedskin, such as fine lines, wrinkles, and sagging skin, in order toimprove the aesthetic appearance of skin by inhibiting C-reactiveprotein secretion or production.

Yet another object of the invention provides a method of detectingCRP-associated inflammation and aging skin by reacting a skin samplewith a probe and detecting the presence of the probe, where the probedetects a CRP and detection of the probe indicates CRP-associatedinflammation and a risk of CRP-associated progression of aging skin.

A further embodiment of the invention relates to a method of monitoringa skin sample by treating the skin sample with a test compositionforming a treated skin sample, reacting the treated skin sample with aprobe, reacting a control skin sample with a probe, and comparing thecontrol skin sample to the treated skin sample for the presence of theprobe, where the presence of the probe identifies the presence of CRP,thereby monitoring the treatment of a skin sample.

These and other objects and advantages of the present invention, andequivalents thereof, are achieved by cosmetic compositions having anatural plant ingredient or blends thereof, and methods of use of suchcompositions for topical application in order to improve the aestheticappearance of skin.

DETAILED DESCRIPTION OF THE INVENTION

In accordance with the foregoing objectives and others detailed herein,the invention overcomes deficiencies associated with the prior art byproviding compositions comprising at least one C-reactive protein (CRP)inhibitor which improves the aesthetic appearance or condition of skinresulting from the chronological aging process. The composition andmethods thereof, once applied to a biological surface or syntheticbiological surface, rejuvenates or enhances the surfaces by providing avariety of anti-inflammatory, anti-aging, and skin texture benefits thatare both visible and invisible. CRP may also be used as a biomarker forCRP-associated inflammation and chronological aging.

In yet another embodiment, the topical compositions having at least oneactive ingredient that acts as a CRP inhibitor may be used to treat,prevent, ameliorate, and/or reduce dermatological signs ofchronologically aging, which represent the structural, functional, andmetabolic changes in the skin that parallel the aging and degenerativechanges in other body organs, or photo-aged skin, which is a separateprocess and largely involves damage to the collagen and elastin fibersin the skin, as fine lines, wrinkles, and sagging skin, in order toimprove the condition and aesthetic appearance of skin.

Another embodiment of the invention relates to compositions having anactive component derived from a natural plant material or combinationsthereof, where a “natural plant material” includes an ingredient,component, constituent, or extract derived from the natural plantmaterial, or synthetic equivalent thereof. The active component inhibitsthe expression, production, or secretion of CRP, and upon application tobiological surfaces, improves the aesthetic appearance or condition ofthe skin by decreasing the natural aging process, inflammation, andchronic and cumulative damage to biological surfaces. Improvements inthe aesthetic appearance of biological surfaces, e.g., skin, may bepreferably achieved by topical application of these compositions to theskin on a daily basis.

A further embodiment of the invention relates to the novel use ofnatural plant materials in the genus Humulus and family Cannabaceae,including but not limited to, H. scandens, H. japonicus, H. lupulus, H.yunnanensis plants, or similar plants, alone or in combination, in atopical cosmetic composition for application on the face, body, and/orhair in order to improve the condition and aesthetic appearance of skinaffected by the natural aging process, where chronic and cumulativedamage to the skin matrix and inflammation normally occur. It is alsocontemplated that other natural plant materials may work equally as wellif they inhibit, reduce, or decrease production, secretion, orexpression of CRP associated with dermatological signs of aging and/ortheir underlying causative factors. In another embodiment of theinvention, any natural plant material or active ingredient that acts asa CRP inhibitor may be useful in inhibiting, reducing, treating,preventing, or reversing the aging process or results therefrom.

For purposes of the invention, the natural plant material may be in anyform including, but not limited to, the whole plant, a dried plant, aground plant, or parts thereof, including but not limited to, seeds,needles, leaves, roots, bark, cones, stems, rhizomes, callus cells,protoplasts, organs and organ systems, and meristems, or componentsand/or constituents found in, or isolated from, the natural plantmaterial, and/or portions of the plant, or extracts derived eitherdirectly or synthetically from the plant, or any combinations thereof. Apreferred natural plant material is in the form of an extract derivedfrom the natural plant material; however, one embodiment of theinvention utilizes a synthetic equivalent of the natural plant materialof interest for use in improving the aesthetic appearance or conditionof skin resulting from the natural aging process. It is to be understoodthat “natural plant material” also includes an ingredient, component,constituent, or extract derived from the natural plant material, orsynthetic equivalent thereof.

One embodiment of the composition of the invention, comprises at leastone natural plant extract which inhibits the expression, production, orsecretion of CRP when applied to a biological surface, such askeratinous tissue. The biological surface may be any surface to whichcosmetics, personal care products, dermatological, and pharmaceuticalcompositions are typically applied, including but not limited to skin,lips, hair, nails, and the like. The composition that is applied tobiological surfaces improves the aesthetic appearance of skin bydecreasing the natural aging process, inflammation, and chronic andcumulative damage to biological surfaces.

The skin is made up of multiple layers of cells that are constantlygoing through self shedding and regeneration once every 30 days inrepeated cycles. The layers may be broadly divided into two sections—thetop epidermis and the underlying dermis. Histological studies of theskin show that a wrinkle is formed following a series of major cellularchanges. During the early phase of aging (from age 35-45), there is agradual and progressive slowing of cellular turnover and regeneration.This results in the skin getting thinner. Skin ages as a result of timeand of cell divisions which may be reversible or irreversible. Agingskin may be induced or accelerated by environmental factors, such as butnot limited to sun exposure, pollutants, toxins, and smoking.

Symptoms of chronological aging include: dry and thin skin, finewrinkles, abnormal blood vessels, age spots, and benign and/or malignantskin tumors. Young skin renews itself more frequently than older skin.The top layers thereby lose more moisture due to the aging process, andolder skin has a drier and more dehydrated appearance. Diminishedproduction of collagen leads to fine wrinkles initially observed aroundthe eyes (commonly known as “crow's feet”), forehead, and othersun-exposed areas. More pronounced effects include furrows at the siteof facial expression lines and sagging folds over the eyelids, neck,jaw, and arms. Within the many small, delicate blood vessels supplyingnutrients to the skin, abnormalities develop. This is particularlyconspicuous over the nose and cheeks. Age spots are pigmentations thatsurfaced as a result of a deregulation of pigment cells in sun-exposedareas. Hyper- and hypo-pigmentation are also a result of the agingprocess.

While genetics play a significant role, the number of wrinkles presentis highly dependent on the amount of sun exposure and otherenvironmental factors. The lines in a “lived-in face,” especially forthose who spend a considerable amount of time outdoors, is a consequencein part of oxidative damage due to overexposure to ultraviolet (UV)sunlight—both UVA (responsible for tanning, wrinkling, and melanoma) andUVB (responsible for sunburn and basal and squamous cell carcinoma). UVlight may further damage skin by increasing the production ofproteolytic enzymes that break down collagen, the connective tissuelocated beneath the dermis.

Reduced nutrition to the epidermis from aging is one factor that causescellular exhaustion and weakness. Without proper nutrition to theepidermis, cellular metabolism of the epidermal cell is slowed.Furthermore, the transportation of certain unwanted byproducts ofcellular metabolism such as free radicals are reduced. The accumulationof such free radicals within the cell may lead to undesirable mutationaldamages in the cell and ultimately cancer.

Embodiments of the invention relate to the discovery that natural plantmaterials having a C-reactive protein (CRP) inhibitor in a compositionor formulation diminish skin lines and wrinkles, as well as relievessagging, or other conditions due to aging. “Inflammaging” is a term thatcorrelates inflammatory changes with age-related diseases. Morespecifically, inflammaging is the up-regulation of the inflammatoryresponse that is age-dependent (Franceschi C, Bonafe M, Valensin S,Olivieri F, De Luca M, Ottaviani E, De Benedictis G. Inflamm-aging. “Anevolutionary perspective on immunosenescence.” Ann N Y Acad Sci.;908:208-218, 2000). Without wishing to be bound by theory, CRPinflammation has been associated with aging skin, reflected by wrinklesand sagging, which are accelerated by chronic inflammation. Thus, byinhibiting or decreasing the production, secretion, or stimulation ofCRP in skin, one may improve the condition and aesthetic appearance ofskin.

In yet another embodiment of the invention, methods of using thecomposition having a CRP inhibitor useful for improving the conditionand aesthetic appearance of skin, particularly matured or maturing skin,by any one of the following means: reducing dermatological signs ofchronological aging, photo-aging, hormonal aging, and/or actinic aging;preventing and/or reducing the appearance of lines and/or wrinkles;reducing the noticeability of facial lines and wrinkles, facial wrinkleson the cheeks, forehead, perpendicular wrinkles between the eyes,horizontal wrinkles above the eyes, and around the mouth, marionettelines, and particularly deep wrinkles or creases; preventing, reducing,and/or diminishing the appearance and/or depth of lines and/or wrinkles;improving the appearance of suborbital lines and/or periorbital lines;reducing the appearance of crow's feet; rejuvenating and/or revitalizingskin, particularly aging skin; reducing skin fragility; preventingand/or reversing of loss of glycosaminoglycans and/or collagen;ameliorating the effects of estrogen imbalance; preventing skin atrophy;preventing, reducing, and/or treating hyperpigmentation; minimizing skindiscoloration; improving skin tone, radiance, clarity and/or tautness;preventing, reducing, and/or ameliorating skin sagging; improving skinfirmness, plumpness, suppleness and/or softness; improving procollagenand/or collagen production; improving skin texture and/or promotingretexturization; improving skin barrier repair and/or function;improving the appearance of skin contours; restoring skin luster and/orbrightness; minimizing dermatological signs of fatigue and/or stress;resisting environmental stress; replenishing ingredients in the skindecreased by aging and/or menopause; improving communication among skincells; increasing cell proliferation and/or multiplication; increasingskin cell metabolism decreased by aging and/or menopause; retardingcellular aging; improving skin moisturization; enhancing skin thickness;increasing skin elasticity and/or resiliency; enhancing exfoliation;improving microcirculation; decreasing and/or preventing celluliteformation; and any combinations thereof.

In another embodiment, the present invention encompasses a method ofimproving the condition and aesthetic appearance of skin, therebyreducing inflammation and other signs of aging, comprising applying toan affected area of skin, a composition containing a natural plantmaterial, or extract therefrom, selected from: H. scandens, H.japonicus, H. lupulus, H. yunnanensis, variations thereof, orcombinations thereof; or any natural plant material, or extracttherefrom, that inhibits expression of C-reactive protein production andsecretion.

One embodiment of the invention relates to methods of applying aneffective amount of a natural plant material, or extract derivedtherefrom, to inhibit expression of C-reactive protein production andsecretion to an affected area of the skin. The composition is preferablytopical and applied once daily and remains on the affected area of theskin, where the affected area of the skin includes, but is not limitedto, the face, neck, legs and thighs, scalp, and overall body.

Another embodiment of the present invention relates to a method ofimproving the condition and aesthetic appearance of skin and comprisesapplying to the skin, or introducing via a directed mode of delivery, acomposition including one or more natural plant materials, or componentsderived therefrom, in an amount effective to improve the aestheticappearance of conditions related to skin, where the natural plantmaterial inhibits expression of C-reactive protein production andsecretion.

In a specific embodiment, the component is in an extract of one or moreof the natural plant materials described in a pharmaceutically,physiologically, cosmetically, and dermatologically-acceptable vehicle,diluent, or carrier, where the composition is topically applied to anaffected area of skin and left to remain on the affected area in anamount effective for treatment for the dermatological signs of aging andimproving the condition and aesthetic appearance of skin.

In a further embodiment, the natural plant material as used herein, alsoincludes “synthetic” extracts, i.e. various combinations of known thenatural plant material components and/or constituents that are combinedto substantially mimic the composition and/or activity of the naturalplant material. Such synthetic extracts are included in the term“natural plant material extract.” Most preferably, the syntheticextracts have substantially the same number of active components as anatural plant material. The correspondence of the numerical incidence ofactives between the synthetic extracts and the natural plant materialmay also be described in terms of “percent commonality.”

Another embodiment relates to a synthetic extract having about 50% ormore commonality to the chemical composition of a natural plantmaterial. For example, the synthetic extract has about 50% or more ofthe active components found in the natural plant material. The chemicalcomposition of the synthetic extract may have about 70% or morecommonality to the chemical composition of the natural plant material. Asynthetic extract may have about 90% or more commonality to the chemicalcomposition of the natural plant material.

For use in the compositions of the invention, the natural plant materialand/or active components are derived directly from the natural plantmaterial, including the entire plant, its seeds, needles, leaves, roots,bark, cones, stems, rhizomes, callus cells, protoplasts, organs andorgan systems, and meristems. The components may be in a pure form, asemi-pure form, or unpurified form. The natural plant material in theform of an extract may be obtained by extracting either the entireplant, or various parts thereof, with 80% ethanol.

Briefly, the extraction method involves washing and extracting the plantmaterial using water, followed by concentrating and filtering. The solidmatter is removed and liquid-liquid butanol/water purification stepsoccur. Butanol (lipophilic impurities, and residual pesticides areremoved. The produce is dried and results in a dry purified extract.

In one embodiment, the extraction method may involve a) pretreating theplant material, which involves washing, then cutting or crushing; b)heat concentrating the cut or crushed plant material from step a); c)extracting the concentrate from step b) with water; d) concentrating theconcentrate from step c); e) filtrating the concentrate of step d) andremoving solid matter; f) purifying with liquid-liquid butanol/water; g)removing butanol, lipophilic impurities, residual pesticides, and thelike; h) drying with or without pressure; and i) resulting in the dryand purified plant material extract. One skilled in the art understandsthat a variety of extraction methods may be used including for example,WO 03/079816; WO 04/014404; and WO 04/014958.

Additionally, an organic solvent extraction method which involveswashing and extracting the plant material using an organic solvent maybe used to extract the plant material. Non-limiting examples of organicsolvents include methanol, ethanol, isopropanol, dichloromethane,chloroform, hexane, xylene, and petroleum ether. An extracting machinemay be used for organic solvent extraction as is well known in thefield.

Organic solvent extraction involves collecting the raw materials fromthe plant that contain the desired constituent(s), such as seeds,needles, leaves, roots, bark, cones, stems, rhizomes, callus cells,protoplasts, organs and organ systems, and meristems. These plantmaterials are ground to small particle sizes, and then put into anextracting machine through an inlet for the raw materials by ameasurable charging machine. The plant raw material is pushed in theextracting machine by a thruster, and slowly moves the plant rawmaterial forward. Organic solvent (e.g., ethanol) may be added into themachine through a solvent inlet at the top of a waste discharge outlet.Due to the difference in gravity and equilibrium, the solvent flowstoward the raw material inlet, soaks the materials and flows out fromthe opposite side of the solvent inlet. Since the plant materials andthe solvent move in opposite directions against each other, the plantmaterials are constantly immersed in a solution that contains alow-concentration of extract. As a result of equilibrium, high yield ofplant constituent(s) may be achieved by continuously extracting theplant material against the low-concentration solution.

An extraction time between about 1-8 hours is suitable, and morepreferably is between about 2-6 hours, and most preferably is betweenabout 3-5 hours. The temperature of extraction is between about 30°C.-90° C., preferably between about 40° C.-70° C., and more preferablybetween about 50° C.-60° C. The collected extract is then fine-filteredto remove debris, and may be used directly, or is concentrated, forexample, by distilling the solvent or by other conventional processing,where the extract may also be provided as a dry powder.

Similarly, aqueous-organic solvent extraction involves initiallycollecting raw materials from a plant containing the desiredalkaloid(s), such as seeds, needles, leaves, roots, bark, cones, stems,rhizomes, callus cells, protoplasts, organs and organ systems, andmeristems of the plant, which are ground into small particle sizes. Theground plant material is soaked in aqueous solution that is acidic oralkaline, depending on the solubility and stability of the desiredextract under acidic or alkaline (basic) conditions. For extractionunder acidic conditions, an

The extract is then collected and fine-filtered to remove debris.Alkaline agents (e.g., ammonia) or acidifying agents (e.g., sulfuricacid) may be added to the extract to neutralize the solution byadjusting the pH, depending on the acidity or alkalinity of thecollected extract. The aqueous extract may be used directly,concentrated or dried. Alternatively, organic solvent may then be addedto the neutralized solution to transfer the extract actives from anaqueous phase to an organic phase. Non-limiting examples of such organicsolvents include, but are not limited to, ethanol, isopropanol, butanol,pentanol, hexanol and xylene. The extract comprising the transferredextract actives dissolved in organic solvent may be used directly, usedas a concentrate, or dried.

Different plants containing different constituents may be mixed andextracted together. This process of mixed extraction may preferably beused for extracting those plants containing constituents having similarsolubility in the solvent used for extraction, such as ethanol. Themixture of extracts may be concentrated and stored in an appropriatesolvent.

In another embodiment, the composition may have an extract derived froma natural plant material or active ingredient that inhibits CRPproduction, secretion, or expression in an amount from about 0.0001% toabout 50%, preferably from about 0.001% to about 20%, more preferablyfrom about 0.01% to about 5%, and most preferably from about 0.3% toabout 1%, based on the total weight of the composition, where thecomposition is useful in improving the condition and aestheticappearance of skin.

The active ingredient or component of a cosmetic, dermatological orpharmaceutical composition is derived from any one, or combinationsthereof, of plants including but not limited to: H. scandens, H.japonicus, H. lupulus, H. yunnanensis, any natural plant material, orsynthetic equivalent thereof, that inhibits or decreases CRP expressionlevels, production, or secretion when the composition is applied to abiological surface. The CRP inhibitor active ingredient which isassociated with dermatological signs of chronologically orhormonally-aged or photo-aged skin, such as fine lines, wrinkles, andsagging skin, is useful in improving the condition and aestheticappearance of skin when applied, preferably topically in a daily mannerto biological surfaces including skin. Without wishing to be bound bytheory, the natural plant extracts and active ingredients therein andthereof, which are CRP inhibitors, exert their effects through amechanism of action involving the inflammatory and immune systemsassociated with aging. CRP found in skin is a sign of cumulative andchronic damage to the skin matrix that causes skin matrix degradation.Topical application of the composition having the CRP inhibitorfacilitates the targeted delivery of the active components without therequirement of an injection or the expertise of a health practitioner.

The active ingredient of the present invention, includes at least oneCRP inhibitor, but may include non-limiting examples of activeingredients useful in treating, preventing, arresting, ameliorating,reducing or diminishing, medical and/or cosmetic conditions associatedwith aging and inflammation of the skin. Such conditions, as usedherein, commonly include, but are not limited to, dermatological aging(chronological aging, hormonal aging and/or actinic aging), dermatitis,skin and hair fragility, hirsutism, rosacea, skin blemishes, sensitiveskin, hyperpigmentation or hypopigmentation, thinning skin, roughness,keratosis, skin atrophy, wrinkles, lines, hyperplasia, fibrosis, and anycombinations thereof. The active components of the present invention mayalso be useful in enhancing the general health, vitality, condition, andaesthetic appearance of the skin.

Topical compositions having the aforementioned active components derivedfrom a natural plant material or synthetic equivalent thereof, where theactive components decrease or inhibit CRP expression, production, orsecretion improve the condition, cosmetic, and/or aesthetic appearanceof skin, particularly of aging and/or inflamed skin.

A further embodiment of the invention encompasses the use of CRP as abiomarker for compounds which improve the condition and appearance ofskin associated with dermatological signs of aging, such as fine lines,wrinkles, and sagging skin. If expression levels of such biomarkers aredecreased in the presence of a composition, where the composition may beused in a cosmetic, dermatological, or pharmaceutical composition of theinvention for improving the condition and appearance of inflamed andaging skin. Preferably the active ingredient of the compositioncomprises a natural plant material, or synthetic equivalent thereof thatinhibits CRP expression, production, or secretion. A variety of methodsfor measuring CRP protein and nucleic acid levels in cells that havebeen exposed to one or more test samples may be performed. A probe maybe used to detect the CRP biomarker. As defined herein, a probe refersto a protein that specifically binds CRP or a fragment thereof, and anucleic acid or oligonucleotide that forms a hybrid structure with asequence in a target region of CRP due to complementarily of at leastone sequence in the probe with a sequence in the CRP target region.

Suitable methods include detection and evaluation of gene activation orexpression of, for example, DNA, RNA, or mRNA. As non-limiting examples,polymerase chain reaction (PCR) assays (e.g., RT-PCR), Northernblotting, in situ hybridization, and other assays as known and practicedin the art may be employed to quantify RNA in cells being assayed fortolerance to a particular treatment (see, e.g., J. O'Connell, 2002,RT-PCR Protocols, Humana Press, Totowa, N.J.; R. Rapley and D. L.Manning, 1998, RNA Isolation and Characterization Protocols, HumanaPress; R. Rapley, 2000, Nucleic Acid Protocols Handbook, Humana Press;all of which are herein incorporated by reference). In accordance withsuch assays, if levels of at least one nucleic acid biomarker isdecreased in the presence of one or more test samples, this isindicative that the substance(s) will decrease, reduce, or amelioratethe dermatological signs of aging.

Nucleic acids may hybridize to each other when at least one strand ofnucleic acid anneals to another nucleic acid strand under definedstringency conditions. As is well known in the art, stringency ofhybridization is determined, e.g., by (a) the temperature at whichhybridization and/or washing is performed, and (b) the ionic strengthand polarity (e.g., formamide) of the hybridization and washingsolutions, as well as other parameters. Hybridization requires that thetwo nucleic acids contain substantially complementary sequences;depending on the stringency of hybridization; however, mismatches may betolerated. The appropriate stringency for hybridizing nucleic acidsdepends on the length of the nucleic acids and the degree ofcomplementarity, variables well known in the art.

Furthermore, in addition to nucleic acid probes, protein probes specificto the C-reactive protein may also be useful in detecting the presenceof CRP-associated inflammation and age progression in biologicalsurfaces, including skin. For example, in Western blots, antibodies thatspecifically bind to CRP, or fragments of CRP, may be used to determinethe presence or absence of CRP in control and test samples. The presenceof CRP may be determined through a variety of tests that are well knownto and commonly practiced by those skilled in the art.

These assays are useful in monitoring the affects after dailyapplication over a period of time of the inventive composition to abiological surface, e.g., skin. If expression levels of the CRPbiomarker are decreased in the presence of one or more test samples,this is indicative that the test sample(s) will decrease, reduce, orameliorate the signs of dermatological aging, especially fine lines,wrinkles, and sagging skin, thereby improving the condition andaesthetic appearance of aging skin.

In accordance with the invention, compositions comprising CRP inhibitorsincluding components from the natural plant material H. scandens, H.japonicus, H. lupulus, H. yunnanensis, or combinations thereof, may beuseful in topically applied formulations, anti-oxidants,anti-inflammatories, sunscreens, cosmetics, including makeup,formulations for reducing dermatological signs of aging, includingwrinkles, fine lines, and sagging skin, topicals, skin penetrationenhancers, and the like. Also in accordance with this invention, theactive ingredient being a CRP inhibitor, for example in a natural plantmaterial, and additional constituents in a composition, may beformulated in a variety of product forms. The compositions may beprepared in targeted delivery systems, e.g. creams, lotions, masks,gels, toners, serums, transdermal patches, and the like, particularlyfor topical administration. Targeted delivery and/or penetrationenhancement may also be achieved by iontophoresis.

The present invention further provides the compositions comprising thenatural plant materials preferably for topical administration or fortargeted delivery without inducing significant irritation. The inventivecompositions are suitable for all skin types, such as sensitive, normal,dry, or oily, preferably sensitive to dry skin, as well as mature skin.In particular embodiments, the compositions may be suitable for dryskin. The compositions are applied to the skin for a period of timesufficient to improve the condition or aesthetic appearance of skin. Thecompositions may be applied topically once, twice, or more daily tobiological surfaces, including but not limited to skin, lips, and hair.The daily application may be applied for a period of one week, twoweeks, four weeks, or more. The preferred composition or formulationcomprising a CRP inhibitor may be applied and left on the affected areaonce daily.

The topical compositions may be formulated into liposomes which maycomprise other additives or substances, and/or which may be modified tomore specifically reach or remain at a site following administration.The compositions of embodiments of the present invention yieldimprovements to the conditions and aesthetic appearance of skin bytreating at least one of the previously described conditions, orcombinations thereof.

As will be appreciated by the practitioner, cosmetic treatmentscomprising compositions containing natural plant materials, includingextracts, components, and/or constituents of the invention may becarried out, for example, by topically applying the cosmetic compositionas described herein according to the routine technique for administeringsuch compositions. The topical cosmetic, dermatological, orpharmaceutical composition preferably is applied once daily for a periodof at least one week, but may include a period of about 2, 4, 8, or 12weeks. The cosmetic composition is preferably applied to the face andneck, but may be applied to any area of skin in need of aestheticimprovement, where the cosmetic composition remains on the affected areaof skin, and preferably not removed or rinsed off the skin. Routine andcommonly practiced techniques encompass the application of creams,lotions, gels, masks, sera, ointments, patches, makeup, makeup-removingmilks, sunscreen compositions, or the like, to the skin. Preferably thecosmetic composition is a topical leave on formulation, where sprayingas a form of application is also envisioned.

Another embodiment of the invention encompasses compositions comprisinga cosmetically, dermatologically, or pharmaceutically acceptableformulation which is suitable for contact with living mammalican tissue,including human tissue, or synthetic equivalents thereof, with virtuallyno adverse physiological effect to the user. Compositions embraced bythis invention can be provided in any cosmetically and/ordermatologically suitable form, preferably as a lotion or cream, butalso in an anhydrous or aqueous base, as well as in a sprayable liquidform. Other suitable cosmetic product forms for the compositions of thisinvention include but are not limited to, for example, an emulsion, acream, a balm, a gloss, a lotion, a mask, a serum, a toner, an ointment,a mousse, a patch, a pomade, a solution, a spray, a wax-based stick, ora towelette. In addition, the compositions contemplated by thisinvention can include one or more compatible cosmetically acceptableadjuvants commonly used and known by the skilled practitioner, such ascolorants, fragrances, emollients, humectants, preservatives, vitamins,chelators, thickeners, perilla oil or perilla seed oil (WO 01/66067 to a“Method of Treating a Skin Condition,” incorporated herewith) and thelike, as well as other botanicals such as aloe, chamomile, and the like,and as further described below.

Also, embraced by the invention are transdermal modes of delivery, suchas patches and the like, with or without a suitable penetrationenhancers. The methods and compositions embodied by the inventionprovide a means by which the natural plant material components may beeffectively administered in a transdermal system. Accordingly, atransdermal means of delivering a composition or formulation (often witha penetration enhancing composition) to the skin is that of thetransdermal patch or a similar device as known and described in the art.Examples of such devices are disclosed in U.S. Pat. Nos. 5,146,846;5,223,262; 4,820,724; 4,379,454; and 4,956,171, all of which areincorporated herein by reference and such descriptions are not meant tobe limiting. The transdermal mode of storing and delivering thecompositions onto the skin, including hair, and forming the activecomposition is convenient and well-suited for the purposes of anembodiment of the present invention. In a preferred method, theapplication is through a sustained release vehicle, e.g., a topicallyapplied sustained released patch. Preferably, when a topical patch isused, the patch is applied to the desired area for extended period oftime. Preferably, the extended period of time is greater than one hour,most preferably the extended period of time is overnight, i.e., when theuser is sleeping.

The natural plant material component(s) of the present invention arepreferably contained in a cosmetically, dermatologically,physiologically, and pharmaceutically acceptable vehicle, medium,diluent or carrier, for use in treating, reducing, ameliorating, orpreventing conditions associated with aging and inflammation inparticular CRP-associated inflammation and aging of biological surfaces.

In an embodiment embracing topical application, the compositions of thisinvention comprise a medium (vehicle, diluent or carrier) that iscompatible with mammalian skin, including hair. The compositions can beformulated as an aqueous phase, an oil phase, alcohol, oraqueous/alcohol-based solutions, ointments, creams, lotions, gels, awax-in-water emulsion, or water-in-oil, oil-in-water, of water-oil-watertriple emulsions having the appearance of a cream or gel,microemulsions, or aerosols.

The aqueous phase is a mixture of one or more water soluble or waterdispersible ingredient, which can be liquid, semi-solid or solid at roomtemperature (25° C.). The vehicle comprises or can be in the form of asuspension, dispersion or solution in water or an aqueous-alcoholicvehicle, which may contain a thickener or gellant. A person skilled inthe art can select the appropriate cosmetic form, the ingredientscontained therein, as well as the method for preparing it, on the basisof the knowledge that the skilled artisan possesses.

In one embodiment, the composition may include an aqueous phase whichmay contain water or a mixture of water and at least one hydrophilicorganic solvent in particular an alcohol, especially a linear orbranched lower monoalcohol containing from 2 to 5 carbon atoms, e.g.,ethanol or propanol; a polyol, e.g., propylene glycol, sorbitol,glycerol, diglycerol, panthenol, or polyethylene glycol, and mixturesthereof. This aqueous phase may represent from about 0.5 weight % toabout 99.99 weight %, based upon the total weight of the composition.

Yet another embodiment when the composition of the invention is in theform of an emulsion, the composition may also optionally comprise asurfactant, preferably in an amount of from about 0.1 weight % to about30 weight %, and in particular, from about 1 weight % to about 20 weight%, based upon the total weight of the composition.

In a further embodiment of the invention, the composition may alsocomprise a thickening polymer such as an amphiphilic polyurethane, apolyacrylic homopolymer or copolymer, a polyester, or ahydrocarbon-based resin.

One embodiment of the invention further relates to a composition of theinvention which may also comprise an oil phase containing oil soluble oroil dispersible ingredients that are liquid at room temperature (25° C.)and/or oily or waxy substances that are solid at room temperature, suchas waxes, semi-solids, gums, and mixtures thereof. This oily phase mayalso contain organic solvents.

Suitable oily materials that are liquid at room temperature, oftenreferred to as oils, include hydrocarbon-based oils of animal originsuch as perhydrosqualene; hydrocarbon-based plant oils such as liquidtriglycerides of fatty acids of 4 to 10 carbon atoms, for instance,heptanoic or octanoic acid triglycerides, or oils such as sunflower oil,corn oil, soybean oil, grapeseed oil, castor oil, avocado oil,caprylic/capric acid triglycerides, jojoba oil; linear or branchedhydrocarbons of mineral or synthetic origin such as liquid paraffins andderivatives thereof, petroleum jelly; synthetic esters and ethers, inparticular esters of fatty alcohols, namely; for example, isopropylmyristate, 2-ethylhexyl palmitate, 2-octyldodecyl stearate, isostearylisostearate; hydroxylated esters such as isostearyl lactate, octylhydroxystearate, octyldodecyl hydroxystearate, heptanoates, octanoatesand decanoates of fatty alcohols; polyol esters such as propylene glycoldioctanoate, neopentyl glycol diheptanoate, diethylene glycoldiisononanoate, and pentaerythritol esters; fatty alcohols containingfrom 12 to 26 carbon atoms such as octyldodecanol, 2-butyloctanol,2-hexyldecanol, 2-undecylpentadecanol, oleyl alcohol; partiallyhydrocarbon-based fluoro oils and/or fluorosilicone oils; silicone oilssuch as volatile or non-volatile, linear or cyclic polydimethylsiloxanes(PDMS) that are liquid or semisolid at room temperature such ascyclomethicones and dimethicones, optionally comprising a phenyl group,for instance phenyl trimethicones, siloxanes, and mixtures thereof.These oils are usually present in an amount of about 0 weight % to about90 weight %, preferably from about 1 weight % to about 80 weight % byweight of the oil phase.

The oil phase of the composition of the invention may also comprise oneor more cosmetically acceptable organic solvents. These solvents arepresent in an amount of about 0 weight % to about 60 weight %,preferably about 1 weight % to about 30 weight %, based on the totalweight of the composition, and may be selected from the group consistingof lipophilic organic solvents, amphiphilic organic solvents andmixtures thereof. Suitable solvents which may be used in the compositionof the invention include acetic acid esters such as methyl, ethyl,butyl, amyl or 2-methoxyethyl acetate; isopropyl acetate; hydrocarbonssuch as toluene, xylene, p-xylene, hexane or heptane; ethers containingat least 3 carbon atoms, and mixtures thereof.

The composition of the invention may further comprise any ingredientconventionally used in the cosmetics field. These ingredients includepreserving agents, aqueous phase thickeners (polysaccharide biopolymers,synthetic polymers) and fatty-phase thickeners, fragrances, hydrophilicand lipophilic active agents, and mixtures thereof. The amounts of thesevarious ingredients are those conventionally used in the cosmetics fieldto achieve their intended purpose, and range typically from about 0.01weight % to about 20 weight %, based upon the total weight of thecomposition. The nature of these ingredients and their amounts must becompatible with the production of the compositions of the invention.

The composition of the invention may also comprise an additionalparticulate phase, typically present in an amount of about 0 weight % toabout 30 weight %, based upon the total weight of the composition,preferably from about 0.05 weight % to about 20 weight %, and which cancomprise pigments and/or pearlescent agents and/or fillers used incosmetic compositions. Suitable inorganic pigments include, but are notlimited to, titanium oxide, zirconium oxide and cerium oxide, as well aszinc oxide, iron oxide, chromium oxide and ferric blue. Suitable organicpigments include, but barium, strontium, calcium, and aluminum lakes andcarbon black. Suitable pearlescent agents include mica coated withtitanium oxide, with iron oxide, or with natural pigment.

Fillers are normally present in an amount of about 0 weight % to about30 weight %, based on the total weight of the composition, preferablyabout 0.5 weight % to about 15 weight %. Suitable fillers include talc,silica, zinc stearate, mica, kaolin, nylon (in particular orgasol)powder, polyethylene powder, Teflon®, starch, boron nitride, copolymermicrospheres such as Expancel® (Nobel Industrie; Sweden), Polytrap® (DowCorning, Inc.; Midland, Mich.), and silicone resin microbeads(Tospearl®; GE Toshiba Silicones; Japan).

The oil phase of the compositions of the invention may comprise one ormore waxes, gums, or mixtures thereof. The waxes includehydrocarbon-based waxes, fluoro waxes and/or silicone waxes and can beof plant, mineral, animal and/or synthetic origin. In particular, thewaxes have a melting point of greater than 25° C., preferably greaterthan 45° C. The compositions of the present invention may contain fromabout 0 weight % to about 20 weight % waxes, based upon the total weightof the composition.

The gums are generally high molecular weight polydimethylsiloxanes(PDMSs), cellulose gums or polysaccharides, and the semi-solid materialsare generally hydrocarbon-based compounds, such as, but not limited to,lanolins and derivatives thereof, or alternatively PDMSs. Thecompositions of the present invention may contain from about 0 weight %to about 20 weight % gums, based upon the total weight of thecomposition, typically from about 0.1 weight % to about 10 weight %.

More particularly, the compositions for topical application can be inthe form of a protective care composition for the skin, preferably forthe face, the neck, the hands, the feet, or other areas of the body.Non-limiting examples include day creams or lotions, night creams orlotions, salves, sunscreen creams, fluid lotions, oils, ointments, gels,masks, body milks, makeup (a foundation), artificial tanningcompositions, depilatories, emulsifiers, patches, or a solid which ispoured or cast as a stick or a dish, for example.

Another particular embodiment of the present invention is directed tothe delivery of the described compositions comprising a natural plantmaterial by targeted delivery systems, for example, liposomes,microspheres (see, e.g., U.S. Pat. No. 5,770,222 to Unger et al.,incorporated herein by reference), and the like, so that the naturalplant material compositions and/or active constituents may more readilyreach and affect the dermal-epidermal junction layer of the area wherethe composition is topically applied, e.g., face or neck, or otheraffected areas of the skin.

In another embodiment, the topical compositions of the present inventionmay also include one or more of the following: a skin penetrationenhancer, an emollient, a skin plumper, an optical diffuser, asunscreen, an exfoliation promoter, and an antioxidant. Details withrespect to these and other suitable cosmetic ingredients can be found inthe International Cosmetic Ingredient Dictionary (ICID) and Handbook,10th Edition (2004), published by the Cosmetic, Toiletry, and FragranceAssociation (CTFA), at pp. 2177-2299, which is herein incorporated byreference in its entirety.

An emollient provides the functional benefits of enhancing skinsmoothness, reducing the appearance of fine lines and coarse wrinkles,and moisturizing. Non-limiting examples include isopropyl myristate,petrolatum, isopropyl lanolate, silicones (e.g., methicone,dimethicone), oils, mineral oils, fatty acid esters, or any mixturesthereof. The emollient is preferably present from about 0.1 weight % toabout 50 weight % of the total weight of the composition.

A skin plumper serves as a collagen enhancer to the skin. An example ofa suitable, and preferred, skin plumper is palmitoyl oligopeptide. Otherskin plumpers are collagen and/or glycosaminoglycan (GAG) enhancingagents. The skin plumper is preferably present from about 0.1 weight %to about 20 weight % of the total weight of the composition.

An optical diffuser is a particle that changes the surface optometricsof skin, resulting in a visual blurring and softening of, for example,lines and wrinkles. Examples of optical diffusers that can be used inthe present invention include, but are not limited to, boron nitride,mica, nylon, polymethylmethacrylate (PMMA), polyurethane powder,sericite, silica, silicone powder, talc, Teflon, titanium dioxide, zincoxide, or any mixtures thereof. The optical diffuser is preferablypresent from about 0.01 weight % to about 20 weight % of the totalweight of the composition.

A sunscreen protects the skin from damaging ultraviolet rays. In anillustrative embodiment of the invention, the sunscreen would provideboth UVA and UVB protection, by using either a single sunscreen or acombination of sunscreens. Among the sunscreens that can be employed inthe present compositions are avobenzone, cinnamic acid derivatives (suchas octylmethoxy cinnamate), octyl salicylate, oxybenzone, titaniumdioxide, zinc oxide, or any mixtures thereof. The sunscreen may bepresent from about 1 weight % to about 30 weight % of the total weightof the composition. The addition of a sunscreen may protect the skinfrom ultraviolet radiation.

The compositions of the invention having sunscreen bring aboutadditional improvements to the aesthetic appearance of skin, includingat least one of the following: minimizes sunburning, minimizes tanning,and reduces redness.

In an embodiment of the invention, compositions may also have one ormore exfoliation promoters. Suitable examples of an exfoliation promoterthat can be used in the present compositions include alpha hydroxy acids(AHA); benzoyl peroxide; beta hydroxy acids; keto acids, such as pyruvicacid, 2-oxopropanoic acid, 2-oxobutanoic acid, and 2-oxopentanoic acid;oxa acids as disclosed in U.S. Pat. Nos. 5,847,003 and 5,834,513 (thedisclosures of which are incorporated herein by reference); salicylicacid; urea; or any mixtures thereof. The preferred exfoliation promotersare 3,6,9-trioxaundecanedioic acid, glycolic acid, lactic acid, or anymixtures thereof. (See also, ICID at p. 2205).

When an embodiment of the invention includes an exfoliation promoter,the composition has about 0.1 weight % to 30 weight %, preferably about1 weight % to about 15 weight % and more preferably about 1 wt % toabout 10 wt %, of the exfoliation promoter based on the total weight ofthe composition.

An antioxidant functions, among other things, to scavenge free radicalsfrom skin to protect the skin from environmental aggressors. Examples ofantioxidants that may be used in the present compositions includecompounds having phenolic hydroxy functions, such as ascorbic acid andits derivatives/esters; beta-carotene; catechins; curcumin; ferulic acidderivatives (e.g. ethyl ferulate, sodium ferulate); gallic acidderivatives (e.g. propyl gallate); lycopene; reductic acid; rosmarinicacid; tannic acid; tetrahydrocurcumin; tocopherol and its derivatives;uric acid; or any mixtures thereof. Other suitable antioxidants arethose that have one or more thiol functions (—SH), in either reduced ornon-reduced form, such as glutathione, lipoic acid, thioglycolic acid,and other sulfhydryl compounds. The antioxidant may be inorganic, suchas bisulfites, metabisulfites, sulfites, or other inorganic salts andacids containing sulfur. Compositions of the present invention may havean antioxidant preferably from about 0.001 wt % to about 10 wt %, andmore preferably from about 0.01 wt % to about 5 wt %, of the totalweight of the composition. (See also, ICID at p. 2184).

In an embodiment of the invention, the composition may also have one ormore of the following cosmetic and pharmaceutical active agents,excipients, ingredients, or adjuvants: anesthetics, antibiotics, e.g.,erythromycins and tetracyclines, salicylic acids, anti-allergenics,antifungals, antiseptics, anti-irritants, anti-inflammatory agents,antimicrobials, analgesics, nitric oxide synthase inhibitors, insectrepellents, self-tanning agents, skin penetration enhancers, skincooling agents, chelating agents, colorants including dyes, lakes andpigments that may be untreated or chemically surface treated to improvewetability or some other property, demulcents, emollients, emulsifiers,fragrances, humectants, lubricants, skin protectants, moisturizers, pHadjusters, preservatives, stabilizers, surfactants, thickeners, filmformers, plasticizers, viscosity modifiers, vitamins, or any mixturesthereof. The amounts of these various substances are those that areconventionally used in the cosmetic or pharmaceutical fields to achievetheir intended purposes, for example, they may constitute from about0.01% to 20% of the total weight of the composition.

Non-limiting examples of active agents for formulating into thecompositions of the invention include those reagents having an effect onthe treatment of wrinkles and/or fine lines, in addition to the activesas described, such as keratolytic agents, i.e., an active agent havingdesquamating, exfoliant, or scrubbing properties, or an active agentwhich can soften the horny layer of the skin. Other examples ofanti-wrinkle or anti-fine line active agents include hydroxy acids andretinoids. These agents can be formulated, for example, in amounts offrom about 0.0001% to 5% by weight relative to the total weight of thecomposition.

Suitable hydroxy acids include, for example, glycolic acid, lactic acid,malic acid, tartaric acid, citric acid, 2-hydroxyalkanoic acid, mandelicacid, salicylic acid and alkyl derivatives thereof, including5-n-octanoylsalicylic acid, 5-n-dodecanoylsalicylic acid,5-n-decanoylsalicylic acid, 5-n-octylsalicylic acid,5-n-heptyloxysalicylic acid, 4-n-heptyloxysalicylic acid and2-hydroxy-3-methylbenzoic acid or alkoxy derivatives thereof, such as2-hydroxy-3-methyoxybenzoic acid.

Emulsifiers are typically present in the compositions of the inventionin an amount of about 0.01 weight % to 30 weight %, by weight andpreferably from about 0.1 weight % to 30 weight % by weight relative tothe total weight of the composition. However, not all compositions willnecessarily include emulsifiers. (See e.g., ICID at p. 2276-2285).

Non-limiting examples of suitable thickening agents include xanthan gum,hydroxypropyl cellulose, hydroxyethyl cellulose, carbomer, gum acacia,Sepigel 305 (available from Seppic Co., France), and clays such asmagnesium aluminum silicate. (See, e.g., ICID at p. 2293-2299).

The topical compositions of the present invention may include, and theirutility can be enhanced by one or more humectants, such as ureas,pyrrolidone carboxylic acids, amino acids, sodium hyaluronates, certainpolyols and other compounds with hygroscopic properties. (See ICID at p.2244).

The general activity and mildness to skin of the present topicalcompositions can also be enhanced by neutralization to pH about 3.5 toabout 7.0, most preferably from pH about 3.7 to about 5.6. Thisneutralization is preferably accomplished with one or more of ammoniumhydroxide, potassium hydroxide, sodium hydroxide, arginine or otheramino acids, and/or triethanolamine.

Exemplary retinoids include, without limitation, retinoic acid (e.g.,all-trans or 13-cis) and derivatives thereof, retinol (Vitamin A) andesters thereof, such as retinol palmitate, retinol acetate and retinolpropionate, and salts thereof.

The natural plant materials, or extracts therefrom, of the presentinvention are preferably contained in a cosmetically or dermatologicallyacceptable vehicle, medium, diluent or carrier. The topical compositioncomprising the natural plant materials may be further formulatedaccording to procedures known in the art to provide cosmeticcompositions such as emulsions, gels, creams, lotions, masks, toners,serums, oils, water-in-oil, oil-in-water, water-oil-water tripleemulsions having the appearance of a cream or gel, microemulsions,ointments, pastes, sticks, cakes, pencils, aerosol, and essences, aswell as other topical cosmetic vehicles. It is also contemplated thattopical compositions of the present invention can be incorporated intodelivery systems such as liposomes and topical patches, tapes, andsprays.

In addition, the compositions may be in the form of vesiculardispersions containing ionic and/or nonionic lipids, as described above.Dosage units suitable for such compositions are formulated according tothe conventional knowledge and techniques used in the art.

In a further embodiment of the invention, the compositions for topicalapplication may be in the form of a protective care composition for theskin, preferably for the face, the neck, the hands, the feet, or otherareas of the body. Non-limiting examples include day creams or lotions,night creams or lotions, sunscreen creams, lotions, or oils, body milks,makeup (a foundation), artificial tanning compositions, depilatories,and patches.

EXAMPLES

The following examples describe specific aspects of the invention toillustrate the invention and provide a description of the presentmethods for those skilled in the art. The examples should not beconstrued as limiting the invention, as the examples merely providespecific methodology useful in the understanding and practice of theinvention and its various aspects.

Example 1

A plant extract of the present invention may be extracted from naturalraw materials by using the methods of aqueous-organic solvent extractionas is well known in the art. One such extraction process is set forthbelow. In another embodiment, plant extracts may be commerciallyobtained, for example from Indena S.p.A. (Milan, Italy).

The following describes a suitable method of preparing a natural plantextract useful for preparing a topical composition of the presentinvention.

An extract was obtained by extracting the whole plant or parts thereof,such as for example, the aerial (i.e., above ground) part of the Humulusscandens plant using an butanol extraction scheme as generally known inthe art. Briefly, the Humulus scandens plant or parts thereof (could befirst manually ground into small particle sizes resulting in a powder)were extracted with water. After concentrating and filtering, the solidmatter was removed. The remaining concentrate was further purified and,butanol, and any impurities, for example, lipophilic impurities andresidual pesticides, were removed. The total concentrated extract wasthen dried, for example by lyophilization, resulting in a dry purifiedextract of H. scandens.

Example 2

Any composition comprising ingredients normally used in formulatingcosmetic foundations with C-reactive protein inhibitors, natural plantextracts having similar properties, or synthetic equivalents thereof maybe used (see, e.g., Table 1).

TABLE 1 INGREDIENTS % Water q.s. 100% Thickener 1.00 Preservative 0.65Butylene glycol 13.00 Pigment 16.00 Sunscreen 2.00 Emulsifier 0.70Polymer 13.00 Other active ingredients 4.70 Humulus scandens 0.3

Example 3 Biopsy Assay

Normal human subjects completed a 3-week clinical study in whichformulas containing the active ingredients were applied daily on theforearm under semi-occlusive conditions, after which a punch biopsy wastaken at each site, the biopsy fixed in formalin, and processed forimmunohistochemistry with a C-Reactive Protein (CRP) antibody (R&DSystems; Minneapolis, Minn.).

Twenty-one (21) normal human subjects were tested on a their forearm,free of topical drugs, cosmetics, and damage to the skin, for 3 weeks (3consecutive rounds of 5×24 hour patches under semi-occlusion; i.e., 5days a week for 3 wks) with 8 different active ingredients. Theapplication dose applied to the skin was 2 mg/cm². After the three weeksapplication clinical study, a 2 millimeter punch biopsy was obtainedfrom each site to which a different application was tested. Twoadditional biopsies were taken from each subject, one from an untreatedsite and another from a vehicle-treated site for controls.

Normal subjects, specifically human, were in good health, age 30-65, notpregnant or planning to get pregnant or nursing, free of skin diseases,diabetes, renal diseases, heart condition, or immunological disorders.The subjects' skin photodamage was mild to moderate as determinedvisually. No subject was under systemic corticosteroids, Accutane™,immunosuppressive drugs, or systemic birth control.

Potentially misleading areas, such as sites of inflammation (needlesite, bruises revealed by histology, etc.) or biopsy edges were avoided.Also, vehicle effects were taken into account, where vehicle effects areany effect due to the solvent (rather than the ingredient) used todissolve the ingredient in order for application to the skin. Each punchbiopsy was fixed in 4% paraformaldehyde. The biopsies were embedded inparaffin and sectioned (5 μm thickness), and stained after rehydration.Sections were stained for C-reactive protein (R&D Systems; polyclonalanti C-reactive protein).

Histological evaluation of the stained biopsies was carried out by twotrained experts in a semi-blind directional difference paradigm.Controls were identified but not the formulas containing the differentactive ingredients. The stained biopsies were separated into threecategories: No improvement, Slight improvement, or Marked improvement.Improvement consisted of a reduction in the amount and intensity ofstaining revealed by the antibody.

The number of panelists showing improvement was determined and apercentage of success derived. Of the 21 human subjects, 52% showedimprovement for in the presence of Humulus scandens: 4 subjects showedmarked improvement, 6 slight improvement, 9 showed no improvement and 2could not be evaluated. The seven other tested ingredients failed. Onthe basis of these numbers as well as the observed magnitude of thenon-quantitative response by the experts, only Humulus scandens wasdetermined to elicit an improvement in a majority of subjects.

The content of all patents, patent applications, published articles,abstracts, books, reference manuals and abstracts, as cited herein arehereby incorporated by reference in their entireties to more fullydescribe the state of the art to which the invention pertains.

It should be understood that the foregoing description is onlyillustrative of the present invention. Various alternatives andmodifications can be devised by those skilled in the art withoutdeparting from the invention. Accordingly, the present invention isintended to embrace all such alternatives, modifications and variationsthat fall within the scope of the appended claims.

1. A composition comprising: (a) a cosmetically, dermatologically,pharmaceutically, or physiologically effective amount of at least oneactive ingredient sufficient to inhibit or decrease C-reactive proteinexpression, production or secretion; and (b) a cosmetically,dermatologically, pharmaceutically, or physiologically acceptablevehicle, wherein said composition improves the condition and/oraesthetic appearance of a biological surface.
 2. The composition ofclaim 1, wherein the active ingredient is derived from H. scandens, H.japonicus, H. lupulus, H. yunnanensis, any combinations thereof, orsynthetic equivalents thereof.
 3. The composition of claim 1, whereinthe active ingredient is present in an amount effective to improve thecondition and/or aesthetic appearance of skin.
 4. The composition ofclaim 1, wherein the composition is applied to skin for a period of timesufficient to improve the condition and/or aesthetic appearance of skin.5. The composition of claim 1, wherein the composition is applied toskin at least once daily for a period of time sufficient to improve thecondition and/or aesthetic appearance of skin.
 6. The composition ofclaim 1 wherein the active ingredient is present in an amount from about0.0001 weight % to about 50 weight %, based upon the total weight of thecomposition.
 7. The composition of claim 1, wherein the activeingredient is present in an amount from about 0.001 weight % to about 20weight %, based upon the total weight of the composition.
 8. Thecomposition of claim 1, wherein the improvement in aesthetic appearancecomprises: reducing dermatological signs of chronological aging,photo-aging, hormonal aging, and/or actinic aging; preventing and/orreducing the appearance of lines and/or wrinkles; reducing thenoticeability of facial lines and wrinkles, facial wrinkles on thecheeks, forehead, perpendicular wrinkles between the eyes, horizontalwrinkles above the eyes, and around the mouth, marionette lines, andparticularly deep wrinkles or creases; preventing, reducing, and/ordiminishing the appearance and/or depth of lines and/or wrinkles;improving the appearance of suborbital lines and/or periorbital lines;reducing the appearance of crow's feet; rejuvenating and/or revitalizingskin, particularly aging skin; reducing skin fragility; preventingand/or reversing of loss of glycosaminoglycans and/or collagen;ameliorating the effects of estrogen imbalance; preventing skin atrophy;preventing, reducing, and/or treating hyperpigmentation; minimizing skindiscoloration; improving skin tone, radiance, clarity and/or tautness;preventing, reducing, and/or ameliorating skin sagging; improving skinfirmness, plumpness, suppleness and/or softness; improving procollagenand/or collagen production; improving skin texture and/or promotingretexturization; improving skin barrier repair and/or function;improving the appearance of skin contours; restoring skin luster and/orbrightness; minimizing dermatological signs of fatigue and/or stress;resisting environmental stress; replenishing ingredients in the skindecreased by aging and/or menopause; improving communication among skincells; increasing cell proliferation and/or multiplication; increasingskin cell metabolism decreased by aging and/or menopause; retardingcellular aging; improving skin moisturization; enhancing skin thickness;increasing skin elasticity and/or resiliency; enhancing exfoliation;improving microcirculation; decreasing and/or preventing celluliteformation; or any combinations thereof.
 9. A composition comprising: (a)a cosmetically, dermatologically, pharmaceutically, or physiologicallyeffective amount of a plant extract derived from H. scandens, Humulusjaponicus, Humulus lupulus, and Humulus yunnanensis, any combinationthereof, or synthetic equivalent thereof; and (b) a cosmetically,dermatologically, pharmaceutically, or physiologically acceptablevehicle, wherein said composition improves the condition and/oraesthetic appearance of a biological surface.
 10. The composition ofclaim 9, wherein said plant extract inhibits or decreases C-reactiveprotein expression, production, or secretion.
 11. The composition ofclaim 9, wherein the plant extract is present in an amount effective toimprove the condition and/or aesthetic appearance of skin.
 12. Thecomposition of claim 9, wherein said composition is applied to skin fora period of time sufficient to improve the aesthetic appearance of skin.13. The composition of claim 9, wherein the composition is applied toskin at least once daily for a period of time sufficient to improve theaesthetic appearance of skin.
 14. The composition of claim 9, whereinthe plant extract is present in an amount from about 0.0001 weight % toabout 50 weight %, based upon the total weight of the composition. 15.The composition of claim 9, wherein the plant extract is present in anamount from about 0.001 weight % to about 20 weight %, based upon thetotal weight of the composition.
 16. The composition of claim 9, whereinthe improvement in aesthetic appearance comprises: reducingdermatological signs of chronological aging, photo-aging, hormonalaging, and/or actinic aging; preventing and/or reducing the appearanceof lines and/or wrinkles; reducing the noticeability of facial lines andwrinkles, facial wrinkles on the cheeks, forehead, perpendicularwrinkles between the eyes, horizontal wrinkles above the eyes, andaround the mouth, marionette lines, and particularly deep wrinkles orcreases; preventing, reducing, and/or diminishing the appearance and/ordepth of lines and/or wrinkles; improving the appearance of suborbitallines and/or periorbital lines; reducing the appearance of crow's feet;rejuvenating and/or revitalizing skin, particularly aging skin; reducingskin fragility; preventing and/or reversing of loss ofglycosaminoglycans and/or collagen; ameliorating the effects of estrogenimbalance; preventing skin atrophy; preventing, reducing, and/ortreating hyperpigmentation; minimizing skin discoloration; improvingskin tone, radiance, clarity and/or tautness; preventing, reducing,and/or ameliorating skin sagging; improving skin firmness, plumpness,suppleness and/or softness; improving procollagen and/or collagenproduction; improving skin texture and/or promoting retexturization;improving skin barrier repair and/or function; improving the appearanceof skin contours; restoring skin luster and/or brightness; minimizingdermatological signs of fatigue and/or stress; resisting environmentalstress; replenishing ingredients in the skin decreased by aging and/ormenopause; improving communication among skin cells; increasing cellproliferation and/or multiplication; increasing skin cell metabolismdecreased by aging and/or menopause; retarding cellular aging; improvingskin moisturization; enhancing skin thickness; increasing skinelasticity and/or resiliency; enhancing exfoliation; improvingmicrocirculation; decreasing and/or preventing cellulite formation; orany combinations thereof.
 17. A method of improving the condition and/oraesthetic appearance of skin comprising topically applying to skin acomposition of claim 1, in an amount effective to improve the conditionand/or aesthetic appearance of skin.
 18. The method of claim 17, whereinthe active ingredient is derived from Humulus scandens, Humulusjaponicus, Humulus lupulus, Humulus yunnanensis, combinations thereof,or synthetic equivalents thereof.
 19. The method of claim 17, whereinsaid applying of the composition occurs for a period of time sufficientto improve the aesthetic appearance of skin.
 20. The method of claim 17,wherein said applying of the composition occurs at least once daily fora period of time sufficient to improve the aesthetic appearance of skin.21. The method of claim 17, wherein the active ingredient is present inan amount from about 0.0001 weight % to about 50 weight %, based uponthe total weight of the composition.
 22. The method of claim 17, whereinthe active ingredient is present in an amount from about 0.001 weight %to about 20 weight %, based upon the total weight of the composition.23. The method of claim 17, wherein the improvement in aestheticappearance comprises: reducing dermatological signs of chronologicalaging, photo-aging, hormonal aging, and/or actinic aging; preventingand/or reducing the appearance of lines and/or wrinkles; reducing thenoticeability of facial lines and wrinkles, facial wrinkles on thecheeks, forehead, perpendicular wrinkles between the eyes, horizontalwrinkles above the eyes, and around the mouth, marionette lines, andparticularly deep wrinkles or creases; preventing, reducing, and/ordiminishing the appearance and/or depth of lines and/or wrinkles;improving the appearance of suborbital lines and/or periorbital lines;reducing the appearance of crow's feet; rejuvenating and/or revitalizingskin, particularly aging skin; reducing skin fragility; preventingand/or reversing of loss of glycosaminoglycans and/or collagen;ameliorating the effects of estrogen imbalance; preventing skin atrophy;preventing, reducing, and/or treating hyperpigmentation; minimizing skindiscoloration; improving skin tone, radiance, clarity and/or tautness;preventing, reducing, and/or ameliorating skin sagging; improving skinfirmness, plumpness, suppleness and/or softness; improving procollagenand/or collagen production; improving skin texture and/or promotingretexturization; improving skin barrier repair and/or function;improving the appearance of skin contours; restoring skin luster and/orbrightness; minimizing dermatological signs of fatigue and/or stress;resisting environmental stress; replenishing ingredients in the skindecreased by aging and/or menopause; improving communication among skincells; increasing cell proliferation and/or multiplication; increasingskin cell metabolism decreased by aging and/or menopause; retardingcellular aging; improving skin moisturization; enhancing skin thickness;increasing skin elasticity and/or resiliency; enhancing exfoliation;improving microcirculation; decreasing and/or preventing celluliteformation; or any combinations thereof.